[Histonet] RE: Biotin free detection of goat primary antibodies

[Linda M Watson]

Hello Everyone,
BioCare Medical makes a great reagent that detects goat primary 
antibodies and is a polymer system. This biotin free system eliminates 
the biotin background problems that is most often enhanced in 
combination with HIER and ABC methods of detection.

Linda

C.M. van der Loos wrote:

>Dear Min-Han,We solved this problem with a 3-step approach: goat primary - rabbit anti-goat IgG - anti-rabbit polymer/HRP. The second step we purchased from Southern Biotech Associates, Birmingham, AL. Dilution of the rabbit anti-goat IgG (human IgG adsorbed) came out at 1:2000-5000 (30 min, RT). Any anti-rabbit polymer/HRP will do. Gayle is fully right with respect to the negative control: find a non-immune goat IgG with a known IgG protein concentration. You need that to calculate the final dilution.Hope this helps a bit.Cheers, ChrisChris van der Loos, PhD
>Dept. of Pathology
>Academic Medical Center M2-230
>Meibergdreef 9
>NL-1105 AZ Amsterdam
>The NetherlandsDate: Tue, 27 May 2008 10:55:59 +0800
>From: "Min-Han Tan" <minhan.tan <@t> gmail.com>
>Subject: [Histonet] Biotin free detection of goat primary antibodies
>To: histonet <@t> lists.utsouthwestern.edu
>
>Dear all,
>
>I would like to enquire - I am using a goat primary antibody currently, and
>my negative control (omitting primary antibody) is showing background
>staining - this is likely a result of biotin.
>
>I'd like to enquire if any one is familiar with biotin free systems that can
>detect goat antibodies. A search of the Dako / Envision product line does
>not yield any results. I am very reluctant to add an avidin-biotin blocking
>step to what is already an overnight incubation.
>
>Thanks!
>
>  
>